【作者】 石和元；

【导师】 王平；

【作者基本信息】 湖北中医药大学 ， 中医基础理论， 2014， 博士

【摘要】 目的：中医元气理论是中医理论体系的重要内容，其取材于哲学元气学说，在《黄帝内经》、《难经》多有阐发，历代医家都十分重视人体元气在防治疾病中的重要作用。重视中医元气理论相关研究是老年病、慢性疾病防治的客观要求，也是实现由医疗向保健和预防转变的需要。本课题从中医元气理论自身发展的脉络出发，梳理中医元气理论的基本概念和理论内涵，对中医元气理论文献及培元固本学术流派进行了考证归纳。并结合课题组前期的研究基础，以老年性痴呆这一老年退化性疾病为切入点，探讨了元气与老年性痴呆发病的关系和培元固本法防治老年性痴呆的机理。以APP/PS1双转基因痴呆小鼠作为动物模型，观察了培元固本中药复方治疗老年性痴呆的作用及其机制。旨在为更深入的中医元气理论研究，以及培元固本治法延缓衰老及防治老年病应用研究等提供理论基础和实验依据方法：1、理论探讨：分析整理有关中医元气的古今文献，归纳中医学对元气基本概念、理论内涵、生成、分布、功能等内容的认识。整理研究中医元气理论形成和发展的学术脉络，以及培元固本学术流派的形成、发展和影响。探讨元气与人体脑部功能的相关性，阐释元气损伤与老年性痴呆发病的关系、以及培元固本防治老年性痴呆的治则治法。本文研究认为中医元气理论具有鲜明的中医特色，具有极强的哲学积淀，内涵丰富、实践性强。历代医家在《黄帝内经》、《难经》有关元气的理论指导下，在长期卓有成效的临床实践的过程中，不断丰富和发展中医元气理论，并形成了培元固本学术流派，为人类延缓衰老、防治老年疾病积累了十分宝贵的经验。2、实验研究：选用APP/PSI双转基因痴呆小鼠模型，以Aβ生成、沉积为靶点，研究培元固本中药复方对AD的作用及机制。Morris水迷宫实验、跳台试验、自主活动分析等方法检测小鼠行为学改变。ELISA法检测AD转基因小鼠海马Aβ含量，Western Blot法观察Aβ主要生成分泌酶和降解酶（BACE1、NEP、IDE）的表达及其上游β淀粉样前体蛋白(APP)的表达，比色法测定小鼠血清和脑组织中GSH-Px，SOD活力和MDA含量。结果：Morris水迷宫实验显示，与空白组比较，模型组小鼠的上平台潜伏期和游泳总路程明显延长(P＜0.01)；与模型组比较，其余各给药组小鼠的上平台潜伏期和游泳总路程明显缩短(P＜0.01，P＜0.05)。与西药多奈哌齐对照组比较，中药Ⅰ组和中药Ⅱ组小鼠的上平台潜伏期和游泳总路程无明显差异（P＞0.05）。与空白组比较，模型组小鼠的穿越平台次数、目标象限时间明显减少及第1次抵原平台时间明显延长(P＜0.01)；与模型组比较，各给药组小鼠的穿越平台次数、目标象限时间明显增加及第1次抵原平台时间明显缩短(P＜0.01，P＜0.05)；与西药多奈哌齐对照组比较，中药Ⅰ组和中药Ⅱ组小鼠目标象限时间及第1次抵原平台时间无明显差异（P＞0.05），穿越平台次数减少(P＜0.05)。与空白组比较，模型组小鼠的潜伏期明显缩短和出错次数明显增加(P＜0.01)；与模型组比较，其余各给药组小鼠的潜伏期明显延长和出错次数明显减少(P＜0.01)；与西药多奈哌齐对照组比较，中药Ⅰ组和中药Ⅱ组小鼠的潜伏期和出错次数无显著差异（P＞0.05），避暗实验等其他行为学实验显示出相似的趋势。ELISA检测结果显示，培元固本中药复方治疗后能够显著减少脑组织内可溶性Aβl-40和可溶性Aβl-42浓度。Western blotting结果显示，培元固本中药复方治疗后可以促进脑部APP降解过程，增加NEP、IDE的表达，而对APP和BACE1的表达量没有影响。结论：1、元气充沛、髓充神旺是脑部功能的物质基础；元气亏虚、髓海失养、元神不藏是老年性痴呆发病的病理基础；元气亏虚、因虚致实、痰浊阻窍是老年性痴呆发病的重要环节；培元固本、化痰醒脑益智是防治老年性痴呆的重要治法。2、培元固本法能够改善痴呆小鼠认知功能和自主活动能力。3、培元固本法治疗老年性痴呆的作用机制与通过增强Aβ降解酶NEP、IDE表达减少海马组织Aβ沉积，促进Aβ降解，减轻Aβ神经毒性有关。更多还原

【Abstract】 Objective：Traditional Chinese medicine Qi theory is an important content of thetheoretical system of traditional Chinese medicine， which is based onthe philosophy ofprimordial qi theory， which mentioned some times inthe "Huang Di Nei Jing" and Nan Jing， and ancient physicians haveattached great importance to the prevention and treatment of diseases ofthe human spirit. it is the objective requirement of senile disease， chronicdisease prevention， that Emphasis on the theory of traditional Chinesemedicine research strength. at the same time， it is also the needs tochange from a medical to health care and prevention. This topic set out fromthe Chinese medicine Qi theory of its own context that developed byitself. has carried on the research and induction of the theory of traditionalChinese medicine literature and vigor vitality and reinforce academic schools.Combining the previous research of the group， take senile dementia anddegenerative diseases of the elderly as the breakthrough point to explore themechanism of strength with the Alzheimer disease relationships and cultivatea solid Prevention Act of senile dementia. With APP/PS1double transgenicmice as an animal model， to observe the Pei yuaneffective herbal tonic yuan Decoction in treating senile dementia and itsmechanism of action. Aimed at providing a theoretical basis and experimentalevidence for further more depth research from theory of TCM study， andvitality injury pathogenesis and elderly population strength loss research， thedevelopment research of the product and the system of health care in peiyuansolid methods， also about the mechanism of experimental study andanti-aging therapies and application of prevention and treatment of geriatric onthe basis of the peiyuan solid. Method：1、 Theoretical discussion：Collate the ancient and modern literature about the strength of Chinesemedicine， summarized the basic concepts of strength， the theoreticalconnotation of Medicine，generation，distribution，cognitive function andother content.Study of the formation and development of traditional Chinesemedicine theory vitality academic context，and the formation of Pei-Yuanconsolidate academic schools，the development and impact. To investigate thecorrelation between Qi and the human brain function，explain the relationshipbetween the strength damage and Alzheimer’s disease pathogenesis， aswell as Peiyuanpreventing senile dementia therapy.This paper considers the Qi theory of TCM has a distinctive strengthcharacteristics of Chinese，with a strong philosophical heritage，richnessin content and with enough practical movements. Ancient physicians enrichedand developed the theory of traditional Chinese medicine，vitality and vigorformed consolidate academic schools with the guidance of Qi theory in theHuangdi Neijing，and in the course of long-term fruitful clinical practice，which has accumulated valuable experience for human delay aging，prevention and treatment of age-related diseases.2、 Experimental study：Using APP/PSI double transgenic mouse model of dementia， with A βgeneration， deposited as target to study the role and mechanism of thetraditional Chinese medicine Nourishing strength Da Bu Yuan Jian which wasused for Reinforcing primordial qi for AD. Using morris water mazetest，step-down test，shuttle box test method，dark avoidance test to checkthe ability of learning and memory in mice. with electron microscopy andCongo red staining to test the effect of Dabu yuan Decoction on ADtransgenic mice pathological injury of hippocampal neurons. ELISA method to detect A β content in AD transgenic mouse hippocampal， WesternBlot method is mainly observed expression of Aβ generation and secretion ofenzymes degrading enzyme (α-secretase enzyme：ADAM-10， β-secretaseenzyme：BACE1， γ-secretase：PS-1， degradation enzyme：NEP， IDE)and its expression of upstream β-amyloid ‘precursor protein (APP)， byimmunohisto chemistry to detect pathological changes in the hippocampal AβAD model mice deposition.Results:Morris water maze test showed that compared with the control group，Mice in model group has longer total swimming distance and platform latencyand which were significantly (P <0.01)；compared with the model group，thetreatment group of mice on the platform latency and swimming distance weresignificantly reduced (P<0.01，P<0.05). Compared with the control group ofWestern medicine in donepezil， Dabu yuan decoct Chinese traditionalmedicine group showed no obvious difference about swimming platformlatency and total distance (P>0.05). Compared with the blank group， modelgroup mice’s number through the platform， the target quadrant timedecreased significantly and the first time reach for the original platform wasobviously prolonged (P<0.01)；compared with the model group，the numberthrough the platform and target quadrant time was significantly increased ineach drug-treated group mice，and the first time against the original platformwas significantly shorter (P<0.01，P<0.05)；Compared with western medicinedonepezil group，Da Bu Yuan Jian mice target quadrant time and the first timeagainst the time of the original platform was not significantly different (P>0.05)， the number of platform cross reducing significantly (P<0.05).Compared with the control group，the latency was significantly shorter and thenumber of errors increased significantly in model mice (P<0.01)；comparedwith the model group，the rest of each treatment group mice’s；latency were significantly delayed and error number decreased significantly (P<0.01)； compared with western medicine donepezil group， no significantdifferences in latency and error-free number of Da Bu Yuan Jian mice (P>0.05)， other behavioral experiments， such as passive avoidance experimentsshowed some similar trends. Pathological observation showed that the numberof senile plaques in hippocampus and areas were reduced in model mice，while after the treatment of western medicine and Da Bu Yuan Jian， the micehippocampus neuropathy reduced. ELISA test results showed that Da Bu YuanJian’s therapy can significantly reduce the soluble Aβl-40and Aβl-42concentration of soluble brain tissue. Western blotting results showed thatafter the treatment of Da Bu Yuan Jian， brain APP degradation waspromoted， increased expression ofNEP、IDE， while expression levels ofBACE1and APP were not affected.Conclusion1、Full of vigor and vitality，pulp filling and Shan fulled is the materialbasis of brain function； Qi deficiency， God not reservoir is theimportant link of senile dementia， and also is the pathological basis of seniledementia. Pei-Yuan consolidate， Xingnaoyizhi is the important therapies forAlzheimer’s prevention and treatment.2、 Peiyuan method can improve the abilities about learning andmemory of dementia mice.3、The mechanism of Peiyuan consolidate method in the treatmentof senile dementia was related to the decrease of hippocampal A β depositionand promotion of the degradation of A β.更多还原